Compositions comprising anti-cd38 antibodies and lenalidomide

ABSTRACT

Disclosed herein are compositions and kits which comprise anti-CD38 antibodies and lenalidomide compounds. Also disclosed are methods for treating cancers, such as multiple myeloma, in subjects with the compositions and kits.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Application No. 61/734,524,filed 7 Dec. 2012, and U.S. Application No. 61/769,247, filed 26 Feb.2013, both of which are herein incorporated by reference in theirentirety.

REFERENCE TO A SEQUENCE LISTING SUBMITTED VIA EFS-WEB

The content of the ASCII text file of the sequence listing named“20130404_034543_001P3_seq” which is 56.7 kb in size was created on 4Apr. 2013 and electronically submitted via EFS-Web herewith theapplication is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The field of the present invention relates to anti-CD38 antibodies,lenalidomide, and cancer treatments.

2. Description of the Related Art

Multiple myeloma (MM) is a B cell malignancy. In MM, abnormal plasmacells accumulate in the bone marrow where they interfere with theproduction of normal cells. Current therapy of MM includesadministration of protcasome inhibitors such as bortezomib,immunomodulatory drugs such as lenalidomide and thalidomide, andchemotherapy such as melphalan and prednisonc. While these agents haveimproved survival in multiple myeloma, invariably resistance becomesproblematic and patients succumb from their illness. Multiple myelomathus remains ultimately fatal, with a median survival of approximately 3to 5 years only.

CD38 is expressed on malignant plasma cells. CD38 is a 45 kD type IItransmembrane glycolprotein with a long C-terminal extracellular domainand a short N-terminal cytoplasmic domain. The CD38 protein is abifunctional ectoenzyme that can catalyze the conversion of NAD⁺ intocyclic ADP-ribose (cADPR) and also hydrolyze cADPR into ADP-ribose. CD38is up-regulated and has been implicated in many hematopoieticmalignancies.

Thus, some proposed MM treatments include the administration ofanti-CD38 antibodies. See, for example, WO 2012/041800; de Weers et al.(2011) J I mmunol 186:1840-1848; and Van der Veer et al. (2011)Haematologica 96(2):284-290. Unfortunately, like various drugs andchemotherapies, not all antibodies are the same and not all antibodiesagainst the same antigen exhibit the same activities.

There is thus a need for new and efficacious treatments for extendingsurvival and improving outcome of treatments of multiple myeloma, andmore generally of blood cancers.

DESCRIPTION OF THE DRAWINGS

Both the foregoing general description and the following detaileddescription are exemplary and explanatory only and are intended toprovide further explanation of the invention as claimed. Theaccompanying drawings are included to provide a further understanding ofthe invention and are incorporated in and constitute part of thisspecification, illustrate several embodiments of the invention, andtogether with the description serve to explain the principles of theinvention.

This invention is further understood by reference to the drawingswherein:

FIG. 1A shows the growth rate of tumors in xenograft models implantedwith H929 cells (H929 models).

FIG. 1B shows the growth rate of tumors in xenograft models implantedwith RPMI8226 cells (RPMI8226 models).

FIG. 2A shows the tumor volume of tumors in RPMI8226 models aftertreatment with the indicated dose of hu38SB19 at the indicated times(arrows).

FIG. 2B shows the body weight of the RPMI8226 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 3A shows the tumor volume of tumors in RPMI8226 models aftertreatment with the indicated dose of lenalidomide at the indicated times(arrows).

FIG. 3B shows the body weight of the RPMI8226 models after treatmentwith the indicated dose of lenalidomide at the indicated times (arrows).

FIG. 4A shows the tumor volume of tumors in RPMI8226 models aftertreatment with the indicated dose of hu38SB19 at the indicated times(top arrows) and the indicated dose of lenalidomide at the indicatedtimes (bottom arrows).

FIG. 4B shows the body weight of the RPMI8226 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (top arrows)and the indicated dose of lenalidomide at the indicated times (bottomarrows).

FIG. 5A shows the tumor volume of tumors in H929 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 5B shows the body weight of the H929 models after treatment withthe indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 6A shows the tumor volume of tumors in H929 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 6B shows the body weight of the H929 models after treatment withthe indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 7A shows the tumor volume of tumors in H929 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 7B shows the body weight of the H929 models after treatment withthe indicated dose of hu38SB19 at the indicated times (arrows).

FIG. 8A shows the tumor volume of tumors in H929 models after treatmentwith the indicated dose of lenalidomide at the indicated times (arrows).

FIG. 8B shows the body weight of the H929 models after treatment withthe indicated dose of lenalidomide at the indicated times (arrows).

FIG. 9A is a graph showing the mean wet tumor weights of the RPMI8226models after the indicated treatment with lenalidomide and/or hu38SB19(mAb).

FIG. 9B is a graph showing the median wet tumor weights of the RPMI8226models after the indicated treatment with lenalidomide and/or hu38SB19(mAb).

FIG. 10A shows the tumor volume of tumors in H929 models after treatmentwith the indicated dose of hu38SB19 at the indicated times (top arrows)and the indicated dose of lenalidomide at the indicated times (bottomarrows).

FIG. 10B shows the body weight of the H929 models after treatment withthe indicated dose of hu38SB19 at the indicated times (top arrows) andthe indicated dose of lenalidomide at the indicated times (bottomarrows).

FIG. 11A is a graph showing the mean wet tumor weights of the H929models after the indicated treatment with lenalidomide and/or hu38SB19(mAb).

FIG. 11B is a graph showing the median wet tumor weights of the H929models after the indicated treatment with lenalidomide and/or hu38SB19(mAb).

FIG. 12 is a graph showing the cell surface density of CD38 in multiplemyeloma cell lines.

FIG. 13 is a graph showing that hu38SB19 inhibits RPMI-8226 tumor growthas a single-agent, i.e., as the sole active ingredient.

FIG. 14 is a graph showing that treatment with both hu38SB19 andlenalidomide inhibits growth of RPMI-8226 tumors.

SUMMARY OF THE INVENTION

In some embodiments, the present invention relates to a method oftreating a cancer in a subject which comprises administering one or moreanti-CD38 antibodies and one or more lenalidomide compounds to thesubject. In some embodiments, the cancer is a hematological malignancy.In some embodiments, the cancer is multiple myeloma. In someembodiments, the cancer is a relapsed multiple myeloma or a refractorymultiple myeloma. In some embodiments, the one or more lenalidomidecompounds is lenalidomide. In some embodiments, the one or moreanti-CD38 antibodies are administered in an effective amount, preferablya synergistic amount. In some embodiments, the one or more anti-CD38antibodies and/or the one or more lenalidomide compounds areadministered in a therapeutically effective amount. In some embodiments,at least one of the one or more anti-CD38 antibodies is capable ofkilling a CD38+ cell by apoptosis, antibody-dependent cell-mediatedcytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). Insome embodiments, the antibody is hu38SB19. In some embodiments, atleast one of the one or more anti-CD38 antibodies comprises one or morecomplementarity-determining region having an amino acid sequenceselected from the group consisting of SEQ ID NOs: 13, 14, 81, 15, 16,17, 18, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 and 36. In some embodiments,at least one of the one or more anti-CD38 antibodies is selected fromthe group consisting of: a) an antibody comprising a heavy chaincomprising three sequential CDRs having amino acid sequences consistingof SEQ ID NOs: 13, 15 and either SEQ ID NO: 14 or SEQ ID NO: 81, and alight chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 16, 17 and 18; b) an antibody comprising aheavy chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 25, 26 and 27, and a light chain comprisingthree sequential CDRs having amino acid sequences consisting of SEQ IDNOs: 28, 29 and 30; c) an antibody comprising a heavy chain comprisingthree sequential CDRs having amino acid sequences consisting of SEQ IDNOs: 1, 2 and 3, and a light chain comprising three sequential CDRshaving amino acid sequences consisting of SEQ ID NOs: 4, 5 and 6; d) anantibody comprising a heavy chain comprising three sequential CDRshaving amino acid sequences consisting of SEQ ID NOs: 7, 8 and 9, and alight chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 10, 11 and 12; e) an antibody comprising aheavy chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 19, 20 and 21, and a light chain comprisingthree sequential CDRs having amino acid sequences consisting of SEQ IDNOs: 22, 23 and 24; and f) an antibody comprising a heavy chaincomprising three sequential CDRs having amino acid sequences consistingof SEQ ID NOs: 31, 32 and 33, and a light chain comprising threesequential CDRs having amino acid sequences consisting of SEQ ID NOs:34, 35 and 36. In some embodiments, the antibody comprises a heavy chainhaving a VH variable region represented by SEQ ID NO: 66, and a lightchain having a VL variable region represented by either SEQ ID NO: 62 orSEQ ID NO: 64. In some embodiments, the antibody comprises a heavy chainhaving a VH variable region represented by SEQ ID NO: 72, and a lightchain having a VL variable region represented by either SEQ ID NO: 68 orSEQ ID NO: 70. In some embodiments, the one or more anti-CD38 antibodiesare administered intravenously. In some embodiments, the one or morelenalidomide compounds are administered orally. In some embodiments, theone or more anti-CD38 antibodies and the one or more lenalidomidecompounds are administered sequentially. In some embodiments, the methodfurther comprises administering a dexamethasone compound, preferablydexamethasone, to the subject. In some embodiments, the dexamethasonecompound is administered orally. In some embodiments, the dexamethasonecompound is administered at a low dose. In some embodiments, the one ormore anti-CD38 antibodies, the one or more lenalidomide compounds, andthe dexamethasone compound are administered sequentially. In someembodiments, the method further comprises administering ananti-coagulation agent to the subject. In some embodiments, theanti-coagulation agent is selected from the group consisting of aspirin,warfarin, and low molecular weight heparin. In some embodiments, the oneor more anti-CD38 antibodies, the one or more lenalidomide compounds,and the anti-coagulation agent are administered sequentially.

In some embodiments, the present invention relates to a compositioncomprising a) at least one anti-CD38 antibody, preferably the antibodyis capable of killing a CD38+ cell by apoptosis, antibody-dependentcell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity(CDC); and b) at least one lenalidomide compound, preferablylenalidomide; and, optionally c) a dexamethasone compound, preferablydexamethasone; and, optionally d) an anti-coagulation agent. In someembodiments, the present invention relates to a composition comprisinga) at least one anti-CD38 antibody; and b) at least one lenalidomidecompound; and, optionally i) a dexamethasone compound; and/or ii) ananti-coagulation agent. In some embodiments, the antibody is capable ofkilling a CD38+ cell by apoptosis, antibody-dependent cell-mediatedcytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). Insome embodiments, the antibody is hu38SB19. In some embodiments, thelenalidomide compound is lenalidomide. In some embodiments, thedexamethasone compound is dexamethasone.

In some embodiments, the present invention is directed to a kitcomprising a) a first composition comprising at least one anti-CD38antibody, preferably the antibody is capable of killing a CD38+ cell byapoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC); and b) a second compositioncomprising at least one lenalidomide compound, preferably lenalidomide.In some embodiments, the compositions in the kit are packaged forsequential administration to a subject. In some embodiments, theantibody is hu38SB19. In some embodiments, the kit further includes adexamethasone compound, preferably dexamethasone, and/or ananti-coagulation agent. In some embodiments, the dexamethasone compoundand/or the anti-coagulation agent are packaged sequential administrationto a subject.

In some embodiments, the present invention is directed to a kitcomprising at least one anti-CD38 antibody capable of killing a CD38+cell by apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC),and complement-dependent cytotoxicity (CDC), packaged together with alabel having one or more messages that the at least one anti-CD38antibody shall be administered in combination with lenalidomide, andoptionally with dexamethasone and/or an anti-coagulation agent. In someembodiments, the antibody is hu38SB19. In some embodiments, the kitfurther includes a dexamethasone compound, preferably dexamethasone,and/or an anti-coagulation agent. In some embodiments, the dexamethasonecompound and/or the anti-coagulation agent are packaged sequentialadministration to a subject.

In some embodiments, the present invention is directed to a combinationof: (i) at least one anti-CD38 antibody, preferably the antibody iscapable of killing a CD38+ cell by apoptosis, antibody-dependentcell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity(CDC); and (ii) at least one lenalidomide compound, preferablylenalidomide; and, optionally (iii) a dexamethasone compound, preferablydexamethasone; and, optionally (iv) an anti-coagulation agent. In someembodiments, the present invention relates to a combination comprisinga) at least one anti-CD38 antibody; and b) at least one lenalidomidecompound; and, optionally i) a dexamethasone compound; and/or ii) ananti-coagulation agent. In some embodiments, the antibody is capable ofkilling a CD38+ cell by apoptosis, antibody-dependent cell-mediatedcytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). Insome embodiments, the antibody is hu38SB19. In some embodiments, thelenalidomide compound is lenalidomide. In some embodiments, thedexamethasone compound is dexamethasone. In some embodiments, thecombination is for sequential use in the treatment of a hematologicalmalignancy, preferably multiple myeloma.

In some embodiments, the present invention is directed to use of (i) atleast one anti-CD38 antibody, preferably the antibody is capable ofkilling a CD38+ cell by apoptosis, antibody-dependent cell-mediatedcytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC); and(ii) at least one lenalidomide compound, preferably lenalidomide; and,optionally (iii) a dexamethasone compound, preferably dexamethasone;and, optionally (iv) an anti-coagulation agent for the treatment of ahematological malignancy, preferably multiple myeloma. In someembodiments, the present invention relates to use of a) at least oneanti-CD38 antibody; and b) at least one lenalidomide compound; and,optionally i) a dexamethasone compound; and/or ii) an anti-coagulationagent for the treatment of a hematological malignancy, preferablymultiple myeloma. In some embodiments, the antibody is capable ofkilling a CD38+ cell by apoptosis, antibody-dependent cell-mediatedcytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC). Insome embodiments, the antibody is hu38SB19. In some embodiments, thelenalidomide compound is lenalidomide. In some embodiments, thedexamethasone compound is dexamethasone.

In some of the various embodiments of the present invention, the subjectto be treated is mammalian. In some of the various embodiments of thepresent invention, the subject to be treated is a test animal such as amouse. In some of the various embodiments of the present invention, thesubject to be treated is human.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to methods of treating a cancer in asubject which comprises administering one or more anti-CD38 antibodiesand one or more lenalidomide jurors will compounds to the subject. Asused herein, “treat” or “treating” means to alleviate symptoms,eliminate the causation of the symptoms either on a temporary orpermanent basis, or to prevent or slow the appearance of symptoms of thenamed disorder or condition. As disclosed herein, the efficacy of alenalidomide compound is considerably improved when administered inconjunction with one or more anti-CD38 antibodies according to thepresent invention. In fact, the administration of one or more anti-CD38antibodies which exhibit (a) the capability of killing a CD38⁺ cell byapoptosis, (b) antibody-dependent cell-mediated cytotoxicity (ADCC), and(c) complement-dependent cytotoxicity (CDC) is believed to considerablyimprove the efficacy of lenalidomide compounds in the treatment ofhematological malignancies, including MM, to a degree that isunexpectedly more than other anti-CD38 antibodies which do not exhibitall three (a)-(c) activities. Therefore, in some embodiments, the one ormore anti-CD38 antibodies are capable of (a) killing a CD38⁺ cell byapoptosis, (b) antibody-dependent cell-mediated cytotoxicity (ADCC), and(c) complement-dependent cytotoxicity (CDC). In some embodiments, theone or more anti-CD38 antibodies and/or the one or more lenalidomidecompounds are administered in a therapeutically effective amount. Asused herein, a “therapeutically effective amount” of a substance refersto an amount of that substance that results in the alleviation of one ormore symptoms, elimination of the causation of the symptoms either on atemporary or permanent basis, and/or the prevention or reduction in theappearance of symptoms of the named disorder or condition in themajority of subjects afflicted with and similarly treated for the nameddisease or disorder.

In some embodiments, the cancer is one in which CD38 is expressed by themalignant cells. In some embodiments, the cancer is a hematologicalmalignancy of the blood, bone marrow, and/or lymph nodes. In someembodiments, the cancer is a blood cancer. Blood cancers includemycloma, lymphoma and leukemia. The blood cancer might, for instance, beselected from the group consisting of multiple myeloma, non-Hodgkin'slymphoma, Hodgkin's lymphoma, hairy cell leukemia, chronic lymphocyticleukemia, chronic myeloid leukemia, acute myeloid leukemia, and acutelymphocytic leukemia. In some embodiments, the cancer is multiplemyeloma (MM). In some embodiments, the cancer is a relapse MM orrefractory MM. As used herein, relapsed MM refers to clinically activeMM after a period of remission and refractory MM refers to progressiveor stable disease while being treated or progressive disease within 3months of the last does of the prior treatment. See Dimopoulos et al.(2010) Eur J Haematology 88:1-15.

In some embodiments, the subject is mammalian, preferably human. In someembodiments, the subject is an adult human, e.g., at least 18 years. Insome embodiments, the subject is in need of treatment for the cancer. Insome embodiments, the subject has been diagnosed as having the cancer.In some embodiments, the cancer is in partial or complete remission,however, the one or more lenalidomide compounds and the one or moreanti-CD38 antibodies are administered to the subject so as to reduce thelikelihood of relapse. In some embodiments, the subject has a Karnofskyperformance status equal or superior to 60%. The Karnofsky status runsfrom 100 to 0, where 100 is “perfect” health and 0 is death (Karnofskyand Burchenal, 1949, “The Clinical Evaluation of Chemotherapeutic Agentsin Cancer.” In: MacLeod C M (Ed), Evaluation of Chemotherapeutic Agents.Columbia Univ Press). In some embodiments, the subject has undergone atleast one or two prior therapies for multiple myeloma, induction therapybeing considered one prior therapy. In some embodiments, the subjectexhibits evidence that either the cancer progressed while the subjectunderwent a prior therapy, or that the subject was refractory to theprior therapy.

In some embodiments, the anti-CD38 antibodies specifically bind CD38. Insome embodiments, the anti-CD38 antibodies are raised against CD38 or anepitope thereof. In some embodiments, the anti-CD38 antibodies aremonoclonal antibodies. In some embodiments, one or more of the anti-CD38antibodies according to the present invention are monoclonal antibodiesas described in WO 2008/047242, which is herein incorporated byreference in its entirety. In some embodiments, one or more of theanti-CD38 antibodies are monoclonal antibodies 38SB13, 38SB18, 38SB19,38SB30, 38SB31, and 38SB39 as described in WO 2008/047242, which isherein incorporated by reference in its entirety. In some embodiments,the one or more anti-CD38 antibodies are capable of killing CD38 cellsby three different cytotoxic mechanisms, induction of apoptosis,antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC).

The term “antibody” is used herein in the broadest sense and includesmonoclonal antibodies (including full length monoclonal antibodies) ofany isotype such as IgG, IgM, IgA, IgD and IgE, polyclonal antibodies,multispecific antibodies, chimeric antibodies, and antibody fragments.As used herein, the prefix “anti-” when in conjunction with an antigen,indicates that the given antibody is reactive with the given antigen. Anantibody reactive with a specific antigen can be generated by syntheticand/or recombinant methods such as selection of libraries of recombinantantibodies in phage or similar vectors, or by immunizing an animal withthe antigen or an antigen-encoding nucleic acid.

A typical IgG antibody is comprised of two identical heavy chains andtwo identical light chains that are joined by disulfide bonds. Eachheavy and light chain contains a constant region and a variable region.Each variable region contains three segments called“complementarity-determining regions” (“CDRs”) or “hypervariableregions”, which are primarily responsible for binding an epitope of anantigen. They are usually referred to as CDR1, CDR2, and CDR3, numberedsequentially from the N-terminus. The more highly conserved portions ofthe variable regions outside of the CDRs are called the “frameworkregions”. As used herein, “V_(H)” or “VH” refers to the variable regionof an immunoglobulin heavy chain of an antibody, including the heavychain of an Fv, scFv, dsFv, Fab, Fab′ or F(ab′)₂ fragment. Reference to“V_(L)” or “VL” refers to the variable region of the immunoglobulinlight chain of an antibody, including the light chain of an Fv, scFv,dsFv, Fab, Fab′ or F(ab′)₂ fragment.

The antibodies according to the present invention may be, e.g., murine,chimeric, and/or humanized antibodies. As used herein, a “chimericantibody” is an antibody in which the constant region, or a portionthereof, is altered, replaced, or exchanged, so that the variable regionis linked to a constant region of a different species, or belonging toanother antibody class or subclass. “Chimeric antibody” also refers toan antibody in which the variable region, or a portion thereof, isaltered, replaced, or exchanged, so that the constant region is linkedto a variable region of a different species, or belonging to anotherantibody class or subclass. Methods for producing chimeric antibodiesare known in the art. See e.g., Morrison, 1985, Science, 229: 1202; Oict al., 1986, BioTechniques, 4: 214; Gillies et al., 1989, J. Immunol.Methods, 125: 191-202; U.S. Pat. Nos. 5,807,715; 4,816,567; and4,816,397, which are incorporated herein by reference in theirentireties. The term “humanized antibody”, as used herein, refers to achimeric antibody which contain minimal sequence derived from non-humanimmunoglobulin. The goal of humanization is a reduction in theimmunogenicity of a xenogenic antibody, such as a murine antibody, forintroduction into a human, while maintaining the full antigen bindingaffinity and specificity of the antibody. Humanized antibodies, orantibodies adapted for non-rejection by other mammals, may be producedusing several technologies such as resurfacing and CDR grafting. As usedherein, the resurfacing technology uses a combination of molecularmodelling, statistical analysis and mutagenesis to alter the non-CDRsurfaces of antibody variable regions to resemble the surfaces of knownantibodies of the target host. The CDR grafting technology involvessubstituting the complementarity determining regions of, for example, amouse antibody, into a human framework domain, e.g., see WO 92/22653.Humanized chimeric antibodies preferably have constant regions andvariable regions other than the complementarity determining regions(CDRs) derived substantially or exclusively from the corresponding humanantibody regions and CDRs derived substantially or exclusively from amammal other than a human.

Strategies and methods for the resurfacing of antibodies, and othermethods for reducing immunogenicity of antibodies within a differenthost, are disclosed in U.S. Pat. No. 5,639,641, which is herebyincorporated in its entirety by reference. Antibodies can be humanizedusing a variety of other techniques including CDR-grafting (EP 0 239400; WO 91/09967; U.S. Pat. Nos. 5,530,101; and 5,585,089), veneering orresurfacing (EP 0 592 106; EP 0 519 596; Padlan E. A., 1991, MolecularImmunology 28(4/5): 489-498; Studnicka G. M. et al., 1994, ProteinEngineering, 7(6): 805-814; Roguska M. A. et al., 1994, PNAS, 91:969-973), chain shuffling (U.S. Pat. No. 5,565,332), and identificationof flexible residues (PCT/US2008/074381). Human antibodies can be madeby a variety of methods known in the art including phage displaymethods. See also U.S. Pat. Nos. 4,444,887, 4,716,111, 5,545,806, and5,814,318; and international patent application publication numbers WO98/46645, WO 98/50433, WO 98/24893, WO 98/16654, WO 96/34096, WO96/33735, and WO 91/10741 (said references incorporated by reference intheir entireties).

In some embodiments, one or more of the anti-CD38 antibodies accordingto the invention are capable of killing a CD38⁺ cell by apoptosis,antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC). In some embodiments, one ormore of the anti-CD38 antibodies according to the invention are capableof killing said CD38⁺ cells by apoptosis even in the absence of stromacells or stroma-derived cytokines. These activities can be assessed asdescribed in WO 2008/047242, which is hereby incorporated by referencein its entirety.

In some embodiments according to the invention, one or more anti-CD38antibodies are selected from the group consisting of 38SB13, 38SB18,38SB19, 38SB30, 38SB31, 38SB39, and antibodies cross-competing with38SB13, 38SB18, 38SB19, 38SB30, 38SB31 or 38SB39. The hybridoma celllines producing the 38SB13, 38SB18, 38SB19, 38SB30, 38SB31, and 38SB39murine anti-CD38 antibodies have been deposited at the American TypeCulture Collection (10801 University Bid, Manassas, Va., 20110-2209,USA), on 21 Jun. 21, 2006, under the deposit numbers PTA-7667, PTA-7669,PTA-7670, PTA-7666, PTA-7668, and PTA-7671, respectively (as describedin WO 2008/047242, which is herein incorporated by reference in itsentirety).

As disclosed herein, references to SEQ ID NOs refers to the sequencesset forth in the Sequence Listing submitted herewith and also as recitedin WO 2008/047242, which is herein incorporated by reference in itsentirety. In some embodiments, the anti-CD38 antibodies according to thepresent invention may, for instance, comprise a heavy chain comprisingthree sequential CDRs having amino acid sequences represented by SEQ IDNOs: 1, 2, and 3, and a light chain comprising three sequential CDRshaving amino acid sequences represented by SEQ ID NOs: 4, 5, and 6. Anexample of such an antibody is the 38SB13 antibody, which comprises aheavy chain having a V_(H) variable region represented by SEQ ID NO: 50,and a light chain having a V_(L) variable region represented by SEQ IDNO: 38.

In some embodiments, the anti-CD38 antibodies according to the presentinvention may, for instance, comprise a heavy chain comprising threesequential CDRs having amino acid sequences represented by SEQ ID NOs:7, 8, and 9, and a light chain comprising three sequential CDRs havingamino acid sequences represented by SEQ ID NOs: 10, 11, and 12. Anexample of such an antibody is the 38SB18 antibody, which comprises aheavy chain having a V_(H) variable region represented by SEQ ID NO: 52and a light chain having a V_(L) variable region represented by SEQ IDNO: 40.

In some embodiments, the anti-CD38 antibodies according to the presentinvention may, for instance, comprise a heavy chain comprising threesequential CDRs having amino acid sequences represented by SEQ ID NO:13, SEQ ID NO: 15 and either SEQ ID NO: 14 or SEQ ID NO: 81, and a lightchain comprising three sequential CDRs having amino acid sequencesrepresented by SEQ ID NOs: 16, 17, and 18. An example of such anantibody is the 38SB19 antibody, which comprises a heavy chain having aV_(H) variable region represented by SEQ ID NO: 54 and a light chainhaving a V_(L) variable region represented by SEQ ID NO: 42. Specificexamples of humanized versions of 38SB19 (hu38SB19) include antibodiescomprising a heavy chain having a V_(H) variable region represented bySEQ ID NO: 66, and a light chain having a V_(L) variable regionrepresented by either SEQ ID NO: 62 or SEQ ID NO: 64. hu38SB19 is ahumanized anti-CD38 antibody currently undergoing clinical evaluation inCD38-positive hematologic malignancies, including multiple myeloma.Previous and current studies demonstrate that the anti-myeloma activityassociated with this agent involve mechanisms of ADCC, and CDC, as wellas novel, direct apoptotic and anti-ADP-ribosyl cyclase activity. SeeMarie-Cécile Wetzel, Céline Nicolazzi, François Vallée, et al. hu38SB19:characterization of a potent phase I humanized anti-CD38 antibody forthe treatment of multiple myeloma and other hematologic malignancies.AACR Annual Meeting 2013, Abstract #4735.

In some embodiments, the anti-CD38 antibodies according to the presentinvention may, for instance, comprise a heavy chain comprising threesequential CDRs having amino acid sequences represented by SEQ ID NOs:19, 20, and 21, and a light chain comprising three sequential CDRshaving amino acid sequences represented by SEQ ID NOs: 22, 23, and 24.An example of such an antibody is the 38SB30 antibody, which comprises aheavy chain having a V_(H) variable region represented by SEQ ID NO: 56and a light chain having a V_(L) variable region represented by SEQ IDNO: 44.

In some embodiments, the anti-CD38 antibodies according to the presentinvention may, for instance, comprise a heavy chain comprising threesequential CDRs having amino acid sequences represented by SEQ ID NOs:25, 26, and 27, and a light chain comprising three sequential CDRshaving amino acid sequences represented by SEQ ID NOs: 28, 29, and 30.An example of such an antibody is the 38SB31 antibody, which comprises aheavy chain having a V_(H) variable region represented by SEQ ID NO: 58and a light chain having a V₁. variable region represented by SEQ ID NO:46. Specific examples of humanized versions of 38SB31 (hu38SB31) includeantibodies comprising a heavy chain having a V_(H) variable regionrepresented by SEQ ID NO: 72, and a light chain having a V_(L) variableregion represented by either SEQ ID NO: 68 or SEQ ID NO: 70.

In some embodiments, the anti-CD38 antibodies according to the presentinvention may, for instance, comprise a heavy chain comprising threesequential CDRs having amino acid sequences represented by SEQ ID NOs:31, 32 and 33, and a light chain comprising three sequential CDRs havingamino acid sequences represented by SEQ ID NOs: 34, 35, and 36. Anexample of such an antibody is the 38SB39 antibody, which comprises aheavy chain having a V_(H) variable region represented by SEQ ID NO: 60and a light chain having a V_(L) variable region represented by SEQ IDNO: 48.

In some embodiments, the anti-CD38 antibodies according to the inventionare humanized antibodies consisting of two identical heavy chains and oftwo identical light chains, wherein each chain consists of one constantregion and of one variable region.

As used herein, a “lenalidomide compound” refers to lenalidomide((RS)-3-(4-amino-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione) andlenalidomide derivatives. As used herein, “lenalidomide derivatives”refers to compounds which have 4-amino-1-oxo-3H-2-isoindolyl, i.e.,

which may or may not be substituted, as part of its structural formula.For example, “lenalidomide derivatives” include those having thefollowing formula:

wherein R1-R8 are each independently H, a halogen, an alkyl, an alkoxy,amino, or an alkylamine, and wherein R5 may additionally be a doublebonded oxygen. In some embodiments, R5 is H. In some embodiments, R8 isH. In some embodiments, both R5 and R8 are H.

In some embodiments, the one or more anti-CD38 antibodies areadministered in an effective amount. As used herein, an effective amountof the one or more anti-CD38 antibodies is an amount which results in anadditive or a synergistic effect with the one or more lenalidomidecompounds. As used herein, a “synergistic amount” is one that results ina synergistic effect. As used herein, a “synergistic effect” refers tothe effect of the combination of the one or more anti-CD38 antibodiesand the one or more lenalidomide compounds which is more than theirexpected additive effect. In some embodiments, the one or more anti-CD38antibodies are administered before, during, and/or after theadministration of the one or more lenalidomide compounds. In someembodiments, the one or more anti-CD38 antibodies and the one or morelenalidomide compounds are co-administered in the form of a singlecomposition, e.g., as a mixture.

Thus, in some embodiments, the present invention is directed tocompositions comprising a mixture of at least one anti-CD38 antibody andat least one lenalidomide compound. In some embodiments, the mixturecomprises the at least one anti-CD38 antibody in an amount that resultsin an additive or a synergistic effect with the at least onelenalidomide compound in a subject when both are administered. In someembodiments, the at least one anti-CD38 antibody in the mixture is onewhich is capable of killing a CD38 cell by apoptosis, antibody-dependentcell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity(CDC); and at least one lenalidomide compound.

For the purposes of the present invention, the methods and compositionsof the present invention are not exclusively limited to those which areobtained by physical association of the anti-CD38 antibodies and thelenalidomide compound, but also to those which permit a separateadministration, which can be simultaneous or spaced out over a period oftime. Thus, in some embodiments, the present invention is directed to afirst composition comprising the one or more anti-CD38 antibodies, and asecond composition comprising one or more lenalidomide compounds. Insome embodiments, the at least one anti-CD38 antibody is one which iscapable of killing a CD38⁺ cell by apoptosis, antibody-dependentcell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity(CDC); and at least one lenalidomide compound. In some embodiments, theamount of the one or more anti-CD38 antibodies provided in the firstcomposition is one that results in an additive or a synergistic effectwith the at least one lenalidomide compound in the second composition ina subject when both are administered.

In some embodiments, the first and second compositions may be packagedin a kit. Thus, in some embodiments, the present invention is directedto kits which comprise a first composition comprising the one or moreanti-CD38 antibodies, and a second composition comprising one or morelenalidomide compounds. In some embodiments, the first and secondcomposition may be mixed together before administering to a subject. Insome embodiments, the first and second compositions, may be administeredeither simultaneously or sequentially (i.e., spaced out over a period oftime) so as to obtain the maximum efficacy, additivity, synergy, or acombination thereof of the combination. In some embodiments, the presentinvention is directed to kits comprising at least one anti-CD38 antibodypackaged together with a label having one or more messages that theanti-CD38 antibody shall or might be administered in combination withlenalidomide and optionally with dexamethasone and/or ananti-coagulation agent. The kits according to the present invention mayfurther comprise one or more messages that the antibody shall or mightbe administered to a subject suffering from a blood cancer such asmultiple myeloma (e.g., relapsed or refractory multiple myeloma). Insome embodiments, the one or more anti-CD38 antibodies in the kits ofthe present invention are those which are capable of killing a CD38 cellby apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC).

In some embodiments, the compositions of the present invention arepharmaceutical compositions. As used herein, the term “pharmaceuticalcomposition” refers to a composition comprising at least one activeprinciple (e.g., an anti-CD38 antibody or a lenalidomide compound) andat least one pharmaceutically acceptable carrier. Pharmaceuticallyacceptable carriers are well known to the skilled in the art, andusually depend on the chosen route of administration. Pharmaceuticalcompositions according to the present invention may be provided in anyform or formulation that is suitable for the chosen route ofadministration, such as e.g., a solution in case of an intravenous routeof administration, e.g., capsules, pills or tablets in case of an oralroute of administration, etc.

The dosage regimen of the active principles and of the pharmaceuticalcomposition described herein can be chosen by prescribing physicians,based on their knowledge of the art, including information published byregulatory authorities. For example, lenalidomide is typicallyadministered orally. According to the European Medicines Agency (EMA),the recommended dose of lenalidomide is 25 mg orally once daily on days1-21 of repeated 28-day cycles. Since, however, co-administration of theone or more anti-CD38 antibodies and the one or more lenalidomidecompounds results in an additive or a synergistic effect, the dosing ofthe lenalidomide compound may be adjusted accordingly, e.g., the dosechanged and/or the dosing schedule modified. Of course, prescribingphysicians might reconsider which dose and schedule to use depending onthe condition and disease status of the patient and based upon clinicaland laboratory findings.

As lenalidomide is approved for the treatment of MM in combination withdexamethasone, the methods and compositions of the present invention mayfurther include dexamethasone, which is member of the glucocorticoidclass of steroid drugs, and acts as an anti-inflammatory andimmunosuppressant. Thus, in some embodiments, the treatment methods ofthe present invention further comprise administering a dexamethasonecompound to the subject being treated with the one or more anti-CD38antibodies and the one or more lenalidomide compounds. Similarly, thecompositions and kits of the present invention which comprise the one ormore anti-CD38 antibodies and/or the one or more lenalidomide compoundsmay further comprise a dexamethasone compound. As used herein, a“dexamethasone compound” refers to dexamethasone((8S,9R,10S,11S,13S,14S,16R,17R)-9-Fluoro-11,17-dihydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one)and dexamethasone derivatives. As used herein, a “dexamethasonederivative” refers to a compound having the following structuralformula:

wherein R1-R17 are each independently H, a halogen, an alkyl, an alkoxy,amino, or an alkylamine. In some preferred embodiments, R1-R3 are H. Insome preferred embodiments, R4-R6 are methyl. In some preferredembodiments, R7 is a halogen, preferably fluorine. In some preferredembodiments, R8 is H. In some preferred embodiments, R1-R3 are H, R4-R6are methyl, R7 is a halogen, preferably fluorine, and R8 is H.

In some embodiments, the dexamethasone compound may be administeredorally. According to the EMA, when combined with lenalidomide, therecommended dose of dexamethasone is 40 mg orally once daily on days1-4, 9-12, and 17-20 of each 28-day cycle for the first 4 cycles oftherapy and then 40 mg once daily on days 1-4 every 28 days. Prescribingphysicians might also re-evaluate which dose of dexamethasone to useupon clinical and laboratory findings.

However, in some embodiments, the dexamethasone compound may beadministered at a lower dose than the dose recommended for dexamethasoneby the EMA. Indeed, recent studies suggest that lenalidomide plus lowdose dexamethasone is associated with better short-term overall survivaland with lower toxicity than lenalidomide plus high-dose dexamethasonein patients with newly diagnosed myeloma (Rajkumar et al. (2010) LancetOnco. 11:29-37). Therefore, in some embodiments of the presentinvention, the dexamethasone compound is administered at low dose. Theterm “low dose” in this context refers to any dose that is at least 20,30 or 40% lower that the dose of dexamethasone recommended by EMA at thedate of first marketing approval of the lenalidomide plus dexamethasonecombination. For instance, administration of 40 mg of dexamethasone ondays 1, 8, 15, and 22 of a 28-day cycle is considered as a low dose ofdexamethasone.

In some embodiments, the methods and compositions of the presentinvention may further include an anti-coagulation agent, such as e.g.,aspirin, warfarin, low molecular weight heparin or equivalentanti-platelet therapeutic. For example, in some embodiments, thetreatment methods of the present invention further compriseadministering an anti-coagulation agent to the subject being treatedwith the one or more anti-CD38 antibodies and the one or morelenalidomide compounds. Similarly, the compositions and kits of thepresent invention which comprise the one or more anti-CD38 antibodiesand/or the one or more lenalidomide compounds may further comprise ananti-coagulation agent.

The compositions of the present invention may be used as a medicamentand/or for use in the manufacture of a medicament. In some embodiments,the compositions of the present invention may be used as a medicamentand/or for use in the manufacture of a medicament for use in thetreatment of a cancer such as a hematological malignancy of the blood,bone marrow, and/or lymph nodes, preferably a blood cancer.

Several documents are cited throughout the text of this specification.Each of the documents herein (including any journal article or abstract,published or unpublished patent application, issued patent,manufacturer's specifications, instructions, etc.) are herebyincorporated by reference. However, there is no admission that anydocument cited herein is indeed prior art in respect of the presentinvention.

The following examples are intended to illustrate but not to limit theinvention.

EXAMPLES

hu38SB19 was provided in solution at 5 mg/ml, stored at 4° C. It wasdiluted into sterile saline in preparation for dosing, stored at 4° C.and used within 10 days of dilution.

Lenolidomide was obtained from AK Scientific Inc. (Mountain View,Calif.) and prepared as a suspension in 1% (w/v) carboxymethyl cellulose(Sigma). Preparation was made using a mortar and pestle to generate aslurry suspension in the vehicle, diluted to the appropriateconcentration, and used for dosing by oral gavage.

Example 1: Effect of the Administration of Both Anti-CD38 Antibody andLenalidomide in a Mice Model of MM

These studies under this Example were done under approval of the UCSFIACUC.

The subcutaneous multiple myeloma (MM) xenograft mouse models wereestablished using H929 and RPMI8226 cell lines. Specifically, 5-6 weekold female Balb/c Scid mice were obtained from Jackson Lab. Mice werehoused for 7-10 days prior to implantation. Mice were housed in adedicated room in the UCSF Mt Zion Animal Barrier Facility. NCI-H929 andRPMI-8226 cells were obtained from the German Collection ofMicroorganisms and Cell Cultures, DSMZ, (Deutsche Sammlung vonMikroorganismen und Zellkulturen), and grown in sterile suspensionculture in T225 flasks as follows: NCI-H929: RPMI1640+20% FBS+4 mML-glutamine+1 mM sodium pyruvate+50 μM mercaptoethanol. RPMI-8226:RPMI1640+10% FBS+4 mM L-glutamine.

At the time of implantation, mice were shaved on the right flank andshoulder region and anesthetized with ip avertin. MM cells suspended inserum free RPMI 1640 media diluted 1:1 with Matrigel (BD) at aconcentration of 1×10⁸ cells per ml were injected sc into the rightflank in 100 μL volume (1×10⁷ cells) using a 1 ml syringe and 25 gneedle. Mice were monitored twice weekly for the appearance of tumorsand once tumors were visible, measurements were collected twice weeklyfor body weight and tumor volume. Electronic balance and calipers wereused and data was collected directly into a study management program(Study Director). When the mean tumor volume reached about 150-200 mm³,the mice were distributed into treatment groups of 8-10 mice per groupsand dosing was begun.

The dosing schedule was hu38SB19 was 2×/wk×2 wk (iv lateral tail vein)and lenalidomide was qd×7×3 wk (po) (orally, one dose per day, 7 days aweek, for 21 days). Dose levels for use in combination studies are asfollows:

Cell Type Lenalidomide hu38SB19 H929 1 mpk 0.5 mpk RPMI8226 15 mpk 15mpk mpk = mg per kg body weight

Data were collected using electronic balance and calipers using a studymanagement application called StudyLog (Study Director). Graphs aretaken directly from the application. The experimental results areprovided in FIGS. 1A-11B.

Based on the single agent results of hu38SB19 and lenalidomide inRPMI-8226 and NCI-H929 multiple myeloma xenograft models, NCI-H929appears to be a more sensitive model to both agents while RPMI-8226seems to be more resistant to the treatments even at the highest dosestested (FIGS. 1-3, 5-8). Therefore in the combination studies, asuboptimal dose for each agent was chosen to evaluate the activity ofthe combination treatment (lenalidomide+hu38SB19) in the NCI-H929 modelwhile higher doses of lenalidomide and hu38SB19 were tested in theRPMI-8226 model.

Antitumor activity was determined according to NCI standards based onthe ratio of the median tumor volume change of the treated/median tumorvolume change of the control×100 (% ΔT/ΔC). Low numerical values forΔT/ΔC describe stronger anti-tumor activity. Anti-tumor activity isdefined as ΔT/ΔC≤40% at minimum. ΔT/ΔC<10% is considered high anti-tumoractivity.

In the RPMI-8226 model, hu38SB19 alone at 15 mg/kg/injection (twice aweek for 2 weeks) was inactive with a % ΔT/ΔC of 44%. Treatment withlenalidomide alone at 15 mg/kg/day (dosed daily for three weeks) wasinactive (61% ΔT/ΔC). The combination of hu38SB19 (15 mg/kg/injection)and lenalidomide (15 mg/kg/day) had higher activity with % T/C of 13%(FIG. 4). The results are summarized in Table 1.

TABLE 1 Anti-tumor efficacy of hu38SB19 in combination with lenalidomideagainst RPMI-8226 multiple myeloma model Schedule of % ΔT/ Dose in mg/kgAdministration ΔC Agent (total dose) IV or PO route (D69) Activity PBS —2x/wk x 2 wk (IV) hu38SB19 15 (60)  2x/wk x 2 wk (IV) 44 InactiveLenalidomide 15 (315) QD x 21 d (PO) 61 Inactive hu38SB19 + 15 (60) +2x/wk x 2 wk (IV) + 13 Active Lenalidomide 15 (315) QD x 21 d (PO) %ΔT/ΔC Median tumor volume change of the treated/Median tumor volumechange of the control x 100, IV = intravenous, PO = oral, d = days, wk =week, QD = once daily, PBS: phosphate buffered saline

In the NCI-H929 model, hu38SB19 alone at 0.5 mg/kg/injection (twice aweek for 2 weeks) was active with a % ΔT/ΔC of 10%. Treatment withlenalidomide alone at 1 mg/kg/day (dosed daily for three weeks) wasactive (20% ATA/C). The combination of hu38SB19 (0.5 mg/kg/injection)and lenalidomide (1 mg/kg/day) had higher activity (tumor regression)with % ΔT/ΔC of −8% (FIG. 10). The results are summarized in Table 2.

TABLE 2 Anti-tumor efficacy of hu38SB19 in combination with lenalidomideagainst NCI-H929 multiple myeloma model Schedule of % ΔT/ Dose in mg/kgAdministration ΔC Agent (total dose) IV or PO route (D69) Activity PBS —2x/wk x 2 wk (IV) hu38SB19 0.5 (2) 2x/wk x 2 wk (IV) 10 ActiveLenalidomide   1 (21) QD x 21 d (PO) 20 Active hu38SB19 + 0.5 (2) +2x/wk x 2 wk (IV) + −8 Highly Lenalidomide 1 (21) QD x 21 d (PO) Active% ΔT/ΔC Median tumor volume change of the treated/Median tumor volumechange of the control x 100, IV = intravenous, PO = oral, d = days, wk =week, QD = once daily, PBS: phosphate buffered saline

In both models, the combination treatment inhibited tumor growth to amuch greater extent than a single agent alone, indicating thecombination of hu38SB19 and lenalidomide blocked tumor cell growththrough potential synergistic mechanisms. Although the molecularmechanisms of action of lenalidomide is still unknown, it is generallybelieved that lenalidomide enhances natural killer cell activity whichis important for antibody dependent cellular cytotoxicity (ADCC) anddirectly induces apoptosis in tumor cells. Hu38SB19 has demonstratedpotent ADCC and direct apoptosis induction activity on tumor cells andthese activities are further enhanced by lenalidomide as evidenced bythe experiments herein.

It has been reported that some CD38 antibodies such as Daratumumab isable to induce apoptosis only after cross-linking with a secondaryantibody without much direct effect by itself. However, in preclinicalstudies, hu38SB19 demonstrated potent direct pro-apoptotic activity ontumor cells without cross-linking. Thus, this unique property ofhu38SB19 may also lead to greater tumor cell killing when in combinationwith lenalidomide compared to other CD38 antibodies combined withlenalidomide.

Example 2: Effect of the Administration of Both Anti-CD38 Antibody andLenalidomide in Humans

A Phase 1b study for evaluating the effects of a treatment with hu38SB19combined with lenalidomide and low dose dexamethasone in patients withrelapsed or refractory multiple myeloma is performed as described below.

The main goals of the Phase 1b study include:

-   -   To determine the efficacy and the maximum tolerated dose;    -   To evaluate the safety, including immunogenicity, of hu38SB19 in        combination with lenalidomide in relapse or refractory multiple        myeloma. The severity, frequency and incidence of all toxicities        is assessed;    -   To evaluate the pharmacokinetics (PK) of hu38SB19 when        administered in combination with lenalidomide and the PK of        lenalidomide in combination with HU38SB19 and dexamethasone.    -   To assess the relationship between clinical (adverse event        and/or tumor response) effects and pharmacologic parameters        (PK/pharmacodynamics), and/or biologic (correlative laboratory)        results;    -   Estimate the activity (response rate) using International        Myeloma Working Group defined response criteria of hu38SB19 plus        lenalidomide and dexamethasone; and    -   To describe overall survival, progression free survival (PFS)        and time to disease progression in patients treated with this        combination.

About 20 to 40 patients may be selected based on the following criteria:The patients are male or female, but must be diagnosed with multiplemycloma and be aged of at least 18 years. For each patient, there is adocumentation of at least 2 prior therapies (induction therapy isconsidered one prior therapy). There is no maximum number of priorregimens and prior bone marrow transplant is acceptable. There is aconfirmed evidence of disease progression from immediately prior MMtherapy or refractory to the immediately prior therapy. Patients mayhave received prior immunomodulatory drugs (IMiDs) (e.g., lenalidomideor thalidomide). Patients are with measurable disease. Patients are witha Karnofsky≥60% performance status. Females of childbearing potentialare included provided they have a negative serum or urine pregnancy testwith a sensitivity of at least 50 mIU/mL within 10 to 4 days and againwithin 24 hours prior to prescribing lenalidomide for Cycle 1(prescriptions must be filled within 7 days as required by RevAssist®)and must either commit to continued abstinence from heterosexualintercourse or begin two acceptable methods of birth control, one highlyeffective method and one additional effective method at the same time,at least 28 days before she starts taking lenalidomide. Females ofchildbearing potential must also agree to ongoing pregnancy testing.Ability to understand the purpose and risks of the study and providesigned and dated informed consent and authorization to use protectedhealth information (in accordance with national and local subjectprivacy regulations). The patient must be able to take aspirin daily asprophylactic anti-coagulation therapy (patients intolerant to aspirinmay use warfarin, low molecular weight heparin or equivalentanti-platelet therapy).

In addition, patients meeting at least one of the following criteria areexcluded:

-   -   Diagnosed or treated for another malignancy within 3 years prior        to enrollment, with the exception of complete resection of basal        cell carcinoma or squamous cell carcinoma of the skin, an in        situ malignancy, or low risk prostate cancer after curative        therapy;    -   Prior anti-cancer therapy (chemotherapy, targeted agents,        radiotherapy, and immunotherapy) within 21 days except for        alkylating agents (e.g., melphalan) where 28 days will be        required or participated in another clinical trial during the        past 30 days;    -   History of significant cardiovascular disease within the past 6        months, unless the disease is well-controlled. Significant        cardiac diseases includes second/third degree heart block;        significant ischemic heart disease (eg, angina); QTc        interval>450 msec at baseline (read by local cardiologist);        poorly controlled hypertension; congestive heart failure of New        York Heart Association (NYHA) Class II (slight limitation of        physical activity;    -   comfortable at rest, but ordinary physical activity results in        fatigue, palpitation, or dyspnea) or worse; left-ventricular        ejection fraction (LVEF)<50%;    -   Prior peripheral stem cell transplant within 12 weeks of the        first dose of study treatment;    -   Daily requirement for corticosteroids (>10 mg/kg prednisone qd)        (except for inhalation corticosteroids);    -   Evidence of mucosal or internal bleeding;    -   Prior radiation therapy or major surgical procedure within 4        weeks of the first dose of study treatment;    -   Known active infection requiring parenteral or oral        anti-infective treatment;    -   Serious psychiatric illness, active alcoholism, or drug        addiction that may hinder or confuse follow-up evaluation;    -   Any medical conditions that, in the Investigator's opinion,        would impose excessive risk to the patient. Examples of such        conditions include any pre-existing kidney disease (acute or        chronic, unless renal insufficiency is felt to be secondary to        MM, hypertension, active seizure disorder or pulmonary diseases        that would impose excessive risk to the patient;    -   Hypersensitivity to any of the components of study therapy that        is not amenable to premedication with steroids and H2 blockers;    -   Known human immunodeficiency virus (HIV) or active hepatitis B        or C viral infection;    -   Neuropathy≥Grade 3 or painful neuropathy≥Grade 2 (National        Cancer Institute Common Terminology Criteria for Adverse Events        [NCI CTCAE] v 4.0);    -   Gastro-intestinal abnormalities, including bowel obstruction,        inability to take oral medication, requirement for        intravenous (IV) alimentation, active peptic ulcer or prior        surgical procedures or bowel resection affecting absorption; and    -   Pregnancy.

The patients are treated with hu38SB19 combined with lenalidomide anddexamethasone. hu38SB19 is administered intravenously as a solution.Lenalidomide is administered orally as capsules. Dexamethasone isadministered orally as tablets. The study duration for an individualpatient includes a screening period for inclusion of up to 21 days, andat least 4 weeks of treatment in the absence of severe adverse reaction,dose limiting toxicity or disease progression plus up to 60 dayspost-treatment follow up. The total duration of the study may be up toone year.

The following parameters are measured during and/or at the end of thestudy:

-   -   Number of patients with adverse events when treated with        hu38SB19 in combination with Lenalidomide    -   Assessment of partial response, complete response, progression        free survival, and survival;    -   Assessment of the following PK parameters: area under curve        (AUC), maximum concentration (Cmax) and plasma half-life (T ½)    -   Number of CD38 receptors occupied by hu38SB19; and    -   Number of anti-SAR antibodies in response to hu38SB19.

Example 3: Efficacy of Anti-CD38 Antibody in In Vivo Tumor Models ofMultiple Myeloma as a Single-Agent or in Combination with andLenalidomide in Humans the Standard-of-Care Immunomodulatory TargetingAgent, Lenalidomide A. Materials and Methods

CD38 Density: CD38 density was determined using anti-CD38-PE Quantibrite(BD Biosciences; Cat. 342371) per the manufacturer's recommendedprotocols.

Reagents & Compounds: hu38SB19 was provided by Sanofi Oncology insolution at 5 mg/ml and stored at 4° C. hu38SB19 was diluted intosterile saline in preparation for dosing and used within 10 days ofdilution. hu38SB19 was administered twice weekly×2 wk IV. Lenolidomide(TC27682) was obtained from AK Scientific Inc. (Mountain View, Calif.)and prepared as a suspension in 1% (w/v) carboxymethyl cellulose(Sigma). Preparation was made using a mortar and pestle to generate aslurry suspension in the vehicle, diluted to the appropriateconcentration, and used for dosing by oral gavage. Lenalidomide wasadministered qd×7×3 wk PO.

Test Animals: 5-6 week old female Balb/c Scid mice were obtained fromJackson Lab. Mice were housed for 7-10 days prior to implantation ofmultiple myeloma (MM) cell lines. Mice were housed in a dedicated roomin the UCSF Mt. Zion Animal Barrier Facility.

Cell culture: RPMI-8226 cells were obtained from the German Collectionof Microorganisms and Cell Cultures, DSMZ, (Deutsche Sammlung vonMikroorganismen und Zellkulturen), and grown in sterile suspensionculture in T225 flasks. RPMI-8226 were cultured in RPMI1640+10% FBS+4 mML-glutamine.

Xenograft Model: At the time of implantation, mice were shaved on theright flank and shoulder. MM cells were suspended in serum free RPMI1640 media diluted 1:1 with Matrigel (BD) at a concentration of 1×10⁸cells per ml were injected sc into the right flank in 100 ul volume(1×10⁷ cells) using a 1 ml syringe and 25 g needle. Mice were monitoredtwice weekly for the appearance of tumors and once tumors were visible,measurements were collected twice weekly for body weight and tumorvolume. Electronic balance and calipers were used and data was collecteddirectly into a study management program (Study Director). When the meantumor volume reached approximately 150-200 mm³, mice were distributedinto treatment groups of 8-10 mice per group and dosing was initiated.

B. Summary and Conclusions

hu38SB19 is a humanized anti-CD38 antibody whose anti-myeloma effectsincorporate mechanisms of ADCC, CDC, and direct apoptosis. FIG. 12 showsthe cell surface density of CD38 in multiple myeloma cell lines. See KimD, Park C Y, Medeiros B C, Weissman I L. CD19-CD45 low/−CD38 high/CD138+plasma cells enrich for human tumorigenic mycloma cells. Leukemia. 2012Dec. 26(12):2530-7. CD38-positive multiple mycloma plasma cellsdemonstrate variable CD38 cell surface densities. All cell lines, withthe exception of XG-6, are reported as CD38-positive. See Bataille R,Jégo G, Robillard N, et al. The phenotype of normal, reactive andmalignant plasma cells. Identification of “many and multiple myelomas”and of new targets for myeloma therapy. Haematologica. 2006 September,91(9):1234-40. Binding of hu38SB19 to CD38 also impinges on the ADPRCenzymatic activity of CD38. In vivo, hu38SB19 demonstrates potentanti-tumor effects in multiple mycloma xenografts, a disease largelycharacterized by neoplastic plasma cells expressing CD38. FIG. 13 showsthat single-agent administration of hu38SB19 results in dose-dependentinhibition of tumor growth in an RPMI-8226 hind-flank model. Themagnitude and significance of tumor growth inhibition at the end of thestudy increased with increased doses of hu38SB19. FIG. 14 shows that acombined regimen of hu38SB19 and Lenalidomide results in significanttumor growth inhibition in an RPMI-8226 xenograft model that is notrobustly sensitive to single-agent therapy with Lenalidomide. These datademonstrate that single-agent hu38SB19 inhibits growth of RPMI-8226tumors and combines with sub-efficacious doses of Lenalidomide toproduce significant inhibition of tumor growth. Taken together, thesedata support further evaluation of hu38SB19, both as a single-agent andin combination with standard-of-care treatment regimens, as a potentialtherapy for the treatment of multiple myeloma.

To the extent necessary to understand or complete the disclosure of thepresent invention, all publications, patents, and patent applicationsmentioned herein are expressly incorporated by reference therein to thesame extent as though each were individually so incorporated.

Having thus described exemplary embodiments of the present invention, itshould be noted by those skilled in the art that the within disclosuresare exemplary only and that various other alternatives, adaptations, andmodifications may be made within the scope of the present invention.Accordingly, the present invention is not limited to the specificembodiments as illustrated herein, but is only limited by the followingclaims.

1: A method of treating a cancer in a subject which comprisesadministering one or more anti-CD38 antibodies and one or morelenalidomide compounds to the subject, wherein malignant cells of thecancer express CD38. 2-3. (canceled) 4: The method of claim 1, whereinthe cancer is a relapsed multiple myeloma or a refractory multiplemyeloma. 5: The method of claim 1, wherein the one or more lenalidomidecompounds is lenalidomide. 6: The method of claim 1, wherein the one ormore anti-CD38 antibodies are administered in an effective amount,preferably a synergistic amount. 7: The method of claim 1, wherein theone or more anti-CD38 antibodies and/or the one or more lenalidomidecompounds are administered in a therapeutically effective amount. 8: Themethod of claim 1, wherein at least one of the one or more anti-CD38antibodies is capable of killing a CD38⁺ cell by apoptosis,antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC).
 9. (canceled) 10: The method ofclaim 1, wherein at least one of the one or more anti-CD38 antibodies isselected from the group consisting of: a) an antibody comprising a heavychain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 13, 15 and either SEQ ID NO: 14 or SEQ ID NO:81, and a light chain comprising three sequential CDRs having amino acidsequences consisting of SEQ ID NOs: 16, 17 and 18; b) an antibodycomprising a heavy chain comprising three sequential CDRs having aminoacid sequences consisting of SEQ ID NOs: 25, 26 and 27, and a lightchain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 28, 29 and 30; c) an antibody comprising aheavy chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 1, 2 and 3, and a light chain comprising threesequential CDRs having amino acid sequences consisting of SEQ ID NOs: 4,5 and 6; d) an antibody comprising a heavy chain comprising threesequential CDRs having amino acid sequences consisting of SEQ ID NOs: 7,8 and 9, and a light chain comprising three sequential CDRs having aminoacid sequences consisting of SEQ ID NOs: 10, 11 and 12; e) an antibodycomprising a heavy chain comprising three sequential CDRs having aminoacid sequences consisting of SEQ ID NOs: 19, 20 and 21, and a lightchain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 22, 23 and 24; and f) an antibody comprising aheavy chain comprising three sequential CDRs having amino acid sequencesconsisting of SEQ ID NOs: 31, 32 and 33, and a light chain comprisingthree sequential CDRs having amino acid sequences consisting of SEQ IDNOs: 34, 35 and
 36. 11: The method of claim 10, wherein said antibodycomprises a heavy chain having a V_(H) variable region represented bySEQ ID NO: 66, and a light chain having a V_(L) variable regionrepresented by either SEQ ID NO: 62 or SEQ ID NO:
 64. 12: The method ofclaim 10, wherein said antibody comprises a heavy chain having a V_(H)variable region represented by SEQ ID NO: 72, and a light chain having aV_(L) variable region represented by either SEQ ID NO: 68 or SEQ ID NO:70. 13: The method of claim 1, wherein the one or more anti-CD38antibodies are administered intravenously. 14: The method of claim 1,wherein the one or more lenalidomide compounds are administered orally.15: The method of claim 1, wherein the one or more anti-CD38 antibodiesand the one or more lenalidomide compounds are administeredsequentially. 16: The method of claim 1, and further comprisingadministering a dexamethasone compound, preferably dexamethasone, to thesubject. 17: The method of claim 16, wherein the dexamethasone compoundis administered orally. 18: The method of claim 16, wherein thedexamethasone compound is administered at a low dose. 19: The method ofclaim 16, wherein the one or more anti-CD38 antibodies, the one or morelenalidomide compounds, and the dexamethasone compound are administeredsequentially. 20: The method of claim 1, and further comprisingadministering an anti-coagulation agent to the subject. 21: The methodof claim 20, wherein the anti-coagulation agent is selected from thegroup consisting of aspirin, warfarin, and low molecular weight heparin.22: The method of claim 20, wherein the one or more anti-CD38antibodies, the one or more lenalidomide compounds, and theanti-coagulation agent are administered sequentially. 23: A compositioncomprising a) at least one anti-CD38 antibody, preferably the antibodyis capable of killing a CD38⁺ cell by apoptosis, antibody-dependentcell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity(CDC); and b) at least one lenalidomide compound, preferablylenalidomide; and, optionally c) a dexamethasone compound, preferablydexamethasone; and, optionally d) an anti-coagulation agent. 24: A kitcomprising a) a first composition comprising at least one anti-CD38antibody, preferably the antibody is capable of killing a CD38⁺ cell byapoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), andcomplement-dependent cytotoxicity (CDC); and b) a second compositioncomprising at least one lenalidomide compound, preferably lenalidomide.25-28. (canceled)